The long-range goal of this Program Project is to develop viral vector-based gene transfer strategies for treating genetic and acquired cardiopulmonary disorders. To this end, a group of investigators with diverse, yet complementary interdisciplinary interests and expertise has established an integrated research effort that is underscored by a common interest inpracticalapplications of gene therapy. A major focus of the Programis the development of improved methods of gene transfer. Four of the components (Subproject 3, Subproject 4, Subproject and Pilot 2) are primarily focused onthe development of vectors suitable for transduction of terminally differentiated cells. These include two stable expression vectors, Adeno-associated virus (AAV, Subproject 3) and lentivirus (subproject 4), as wellas two transient expression vectors, adenovirus (Ad, Subproject 5) and the entomopoxvirus (EPV, Pilot 2). In each case, the efforts are directed at identifying and circumventing obstacles to the practical applicationof gene therapy, most of which are rooted in the basic biology of the individual virus upon which each vector system is based. Along with the vector development components, there is a strong emphasis on the development of new animal models, including the development of knock-out mouse models for monogenic cardiac (Subproject 1) and pulmonary (Pilot 1, Subproject 2) disorders, and a transgenic model for AAV integration (Subproject 1). The utilization of established models of hypertension and acute lung injury are likewise pivotal for Subprojects 6 and 5, respectively. The program also will investigate the applications of these vector systems to three cardiopulmonary disorders: alpha-1-antitrypsin deficiency (alpha 1AT, Subproject 2), acid maltase deficiency (GAA, Subproject 1) and hypertension (Subproject 6). This will be done by targeting vectors to muscle (Subprojects 1 and 2) and vascular endothelium (Subprogject 6). Additionally, the Program has established a Vector Core Laboratory (Core B) which will supply vectors of uniform and reproducible quality to all subprojects, and investigate improved methods of generating recombinant AAV (rAAV) and adenovirus (Ad) vectors. The Vector Core will also serve as a mechanism to insure rapid exchange of information among all subprojects. Finally, an Administrative Core (Core A) will insure centralized fiscal management and oversight for the subprojects and pilot/feasibility projects.